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丙酮丁醇梭菌和氨基戊酸梭菌这两种专性厌氧菌对氧应激的适应性反应。

Adaptive responses to oxygen stress in obligatory anaerobes Clostridium acetobutylicum and Clostridium aminovalericum.

作者信息

Kawasaki Shinji, Watamura Yusuke, Ono Masaki, Watanabe Toshihiro, Takeda Kouji, Niimura Youichi

机构信息

Department of Bioscience, Tokyo University of Agriculture, 1-1-1 Setagaya-ku, Tokyo 156-8502, Japan.

出版信息

Appl Environ Microbiol. 2005 Dec;71(12):8442-50. doi: 10.1128/AEM.71.12.8442-8450.2005.

Abstract

Clostridium acetobutylicum and Clostridium aminovalericum, both obligatory anaerobes, grow normally after growth conditions are changed from anoxic to microoxic, where the cells consume oxygen proficiently. In C. aminovalericum, a gene encoding a previously characterized H2O-forming NADH oxidase, designated noxA, was cloned and sequenced. The expression of noxA was strongly upregulated within 10 min after the growth conditions were altered to a microoxic state, indicating that C. aminovalericum NoxA is involved in oxygen metabolism. In C. acetobutylicum, genes suggested to be involved in oxygen metabolism and genes for reactive oxygen species (ROS) scavenging were chosen from the genome database. Although no clear orthologue of C. aminovalericum NoxA was found, Northern blot analysis identified many O2-responsive genes (e.g., a gene cluster [CAC2448 to CAC2452] encoding an NADH rubredoxin oxidoreductase-A-type flavoprotein-desulfoferrodoxin homologue-MerR family-like protein-flavodoxin, an operon [CAC1547 to CAC1549] encoding a thioredoxin-thioredoxin reductase-glutathione peroxidase-like protein, an operon [CAC1570 and CAC1571] encoding two glutathione peroxidase-like proteins, and genes encoding thiol peroxidase, bacterioferritin comigratory proteins, and superoxide dismutase) whose expression was quickly and synchronously upregulated within 10 min after flushing with 5% O2. The corresponding enzyme activities, such as NAD(P)H-dependent peroxide (H2O2 and alkyl hydroperoxides) reductase, were highly induced, indicating that microoxic growth of C. acetobutylicum is associated with the expression of a number of genes for oxygen metabolism and ROS scavenging.

摘要

丙酮丁醇梭菌和氨基戊酸梭菌均为专性厌氧菌,在生长条件从缺氧变为微氧后能正常生长,在此微氧条件下细胞能高效消耗氧气。在氨基戊酸梭菌中,一个编码先前已鉴定的产H₂O的NADH氧化酶(命名为NoxA)的基因被克隆并测序。在生长条件改变为微氧状态后的10分钟内,noxA的表达被强烈上调,这表明氨基戊酸梭菌NoxA参与氧代谢。在丙酮丁醇梭菌中,从基因组数据库中选取了推测参与氧代谢的基因以及活性氧(ROS)清除相关基因。虽然未发现与氨基戊酸梭菌NoxA明显同源的基因,但Northern印迹分析鉴定出许多对O₂有反应的基因(例如,一个编码NADH红素氧还蛋白氧化还原酶 - A型黄素蛋白 - 脱硫铁氧还蛋白同源物 - MerR家族样蛋白 - 黄素氧还蛋白的基因簇 [CAC2448至CAC2452]、一个编码硫氧还蛋白 - 硫氧还蛋白还原酶 - 谷胱甘肽过氧化物酶样蛋白的操纵子 [CAC1547至CAC1549]、一个编码两个谷胱甘肽过氧化物酶样蛋白的操纵子 [CAC1570和CAC1571],以及编码硫醇过氧化物酶、细菌铁蛋白共迁移蛋白和超氧化物歧化酶的基因),其表达在通入5% O₂后10分钟内迅速且同步上调。相应的酶活性,如NAD(P)H依赖性过氧化物(H₂O₂和烷基氢过氧化物)还原酶,被高度诱导,这表明丙酮丁醇梭菌的微氧生长与许多氧代谢和ROS清除相关基因的表达有关。

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